Conflicts of Interest in the Assessment of Chemicals, Waste, and Pollution.

Pollution by chemicals and waste impacts human and ecosystem health on regional, national, and global scales, resulting, together with climate change and biodiversity loss, in a triple planetary crisis. Consequently, in 2022, countries agreed to establish an intergovernmental science-policy panel (SPP) on chemicals, waste, and pollution prevention, complementary to the existing intergovernmental science-policy bodies on climate change and biodiversity. To ensure the SPP's success, it is imperative to protect it from conflicts of interest (COI). Here, we (i) define and review the implications of COI, and its relevance for the management of chemicals, waste, and pollution; (ii) summarize established tactics to manufacture doubt in favor of vested interests, i.e., to counter scientific evidence and/or to promote misleading narratives favorable to financial interests; and (iii) illustrate these with selected examples. This analysis leads to a review of arguments for and against chemical industry representation in the SPP's work. We further (iv) rebut an assertion voiced by some that the chemical industry should be directly involved in the panel's work because it possesses data on chemicals essential for the panel's activities. Finally, (v) we present steps that should be taken to prevent the detrimental impacts of COI in the work of the SPP. In particular, we propose to include an independent auditor's role in the SPP to ensure that participation and processes follow clear COI rules. Among others, the auditor should evaluate the content of the assessments produced to ensure unbiased representation of information that underpins the SPP's activities.

The Role of Behavioral Ecotoxicology in Environmental Protection.

For decades, we have known that chemicals affect human and wildlife behavior. Moreover, due to recent technological and computational advances, scientists are now increasingly aware that a wide variety of contaminants and other environmental stressors adversely affect organismal behavior and subsequent ecological outcomes in terrestrial and aquatic ecosystems. There is also a groundswell of concern that regulatory ecotoxicology does not adequately consider behavior, primarily due to a lack of standardized toxicity methods. This has, in turn, led to the exclusion of many behavioral ecotoxicology studies from chemical risk assessments. To improve understanding of the challenges and opportunities for behavioral ecotoxicology within regulatory toxicology/risk assessment, a unique workshop with international representatives from the fields of behavioral ecology, ecotoxicology, regulatory (eco)toxicology, neurotoxicology, test standardization, and risk assessment resulted in the formation of consensus perspectives and recommendations, which promise to serve as a roadmap to advance interfaces among the basic and translational sciences, and regulatory practices.

Feeding Behavioural Studies with Freshwater Gammarus spp.: The Importance of a Standardised Methodology.

Freshwater Gammarids are common leaf-shredding detritivores, and they usually feed on naturally conditioned organic material, in other words leaf litter that is characterised by an increased palatability, due to the action and presence of microorganisms (Chaumot et al. 2015; Cummins 1974: Maltby et al. 2002). Gammarus spp. are biologically omnivorous organisms, so they are involved in shredding leaf litter and are also prone to cannibalism, predation behaviour (Kelly et al. 2002) and coprophagy when juveniles (McCahon and Pascoe 1988). Gammarus spp. is a keystone species (Woodward et al. 2008), and it plays an important role in the decomposition of organic matter (Alonso et al. 2009; Bundschuh et al. 2013) and is also a noteworthy prey for fish and birds (Andrén and Eriksson Wiklund 2013; Blarer and Burkhardt-Holm 2016). Gammarids are considered to be fairly sensitive to different contaminants (Ashauer et al. 2010; Bloor et al. 2005; Felten et al. 2008a; Lahive et al. 2015; Kunz et al. 2010); in fact Amphipods have been reported to be one of the most sensitive orders to metals and organic compounds (Wogram and Liess 2001), which makes them representative test organisms for ecotoxicological studies and valid sentinel species for assessing water quality status (Garcia-Galan et al. 2017).

Endocrine Disruption in Invertebrates: A Survey of Research Progress.

Manmade chemicals can interfere with endocrine processes and have permeated many ecosystems. Arguably, the most devastating example of endocrine disruption occurred in gastropod molluscs which led to the banning of tributyltin. The invertebrates consist of ∼95% of all known animals and possess endocrine systems that can significantly differ from that of vertebrates. An expert group in the late 1990s highlighted considerable paucity in our knowledge of these endocrine systems and the limited ability to ascertain risks of endocrine-disrupting chemicals (EDCs) to invertebrates. Twenty years later, we surveyed experts in this field on the current state of the science. Respondents agreed that endocrine disruption is still a significant issue and noted that there was key evidence that EDCs were impacting invertebrates groups. Respondents noted a variety of impediments to advancing the science, including inadequate funding, insufficient knowledge to develop appropriate assays, and generally low support for invertebrate studies. Several scientists highlighted that resources were being misdirected with studies that address impacts of vertebrate EDCs or using biomarkers specific to vertebrate endocrine disruption. Sadly, many of the recommendations proposed by respondents matched those made over two decades ago. Accordingly, the field has not advanced as much as one might have expected.

Effects of the antidepressant fluoxetine on pigment dispersion in chromatophores of the common sand shrimp, Crangon crangon: repeated experiments paint an inconclusive picture.

The effects of antidepressants in the environment are starting to generate considerable interest due to the fact that neurotransmitters influence a range of biological processes. Crypsis is an important behavioural and physiological response in many crustaceans modulated by monoamine and pigment dispersing/concentrating hormones. This study aimed to develop a test methodology and investigate the effects of the selective serotonin reuptake inhibitor (SSRI), fluoxetine, on a chromatophore index and overall carapace 'darkness' in the common sand shrimp Crangon crangon. Adult shrimp were exposed for either 1 h, 1 day or 1 week across a range of nominal fluoxetine concentrations (10 ng/L, 100 ng/L and 1000 ng/L) and the chromatophore index or carapace percentage 'darkness' was recorded following 30 min on white and black substrates. These experiments were repeated three times using different specimens. Animals became significantly darker (~20%) on darker background and lighter on light backgrounds as one might expect. However, time periods over which the animals were recorded had a significant impact on the colouration suggesting habituation to laboratory conditions. Fluoxetine exposure came up as a significant factor in two of the three trials for the chromatophore index but the results was inconsistent between trials. There was a high degree of correlation between the chromatophore index and the percentage darkness analyses however, there was no significant effects for fluoxetine exposure with the percentage darkness data. We conclude that the effects on antidepressants on colour change remain inconclusive from these experiments and we discuss potential areas to improve the repeatability of the experiments.

Impacts of ionising radiation on sperm quality, DNA integrity and post-fertilisation development in marine and freshwater crustaceans.

Crustaceans have been designated as internationally important model organisms in the development of environmental radioprotection measures. Despite the known sensitivity of sperm to ionizing radiation, the impacts of chronic radiation exposure on male fertility in crustaceans have not been studied. For the first time, the present study aimed to assess the impacts of chronic radiation exposure on male fertility, sperm DNA damage and concomitant impacts on breeding in two amphipod crustaceans. Echinogammarus marinus and Gammarus pulex (male fertility only) were exposed to phosphorus-32 at dose rates of 0, 0.1, 1 and 10 mGy/d and sperm parameters, DNA damage and knock-on impacts on breeding were assessed. Sperm quality parameters and DNA damage were assessed using a fluorescent staining method and single cell gel electrophoresis respectively. Concomitant effects of male exposure to radiation on fecundity were determined by pairing phosphorus-32 exposed males to unexposed sexually mature females. In E. marinus, a statistically significant reduction of 9 and 11% in the quality of sperm was recorded at dose rates of 1 and 10 mGy/d respectively, with no significant effects recorded on sperm counts. Conversely in the freshwater G. pulex, no significant impact of radiation on sperm quantity or quality was recorded. For E. marinus, a statistically significant increase in DNA damage was recorded at doses of 10 mGy/d. Reduced fecundity and an increase in the frequency of abnormal embryos was recorded in female E. marinus breeding with males exposed to radiation. These findings suggest sperm quality may be a sensitive indicator of radiation exposure in invertebrates with potential impacts on the unexposed embryo, though unclear dose-response and differences between two closely related species necessitate further study before robust conclusions can be drawn.

The effects of fluoxetine on attachment and righting behaviours in marine (Gibbula unbilicalis) and freshwater (Lymnea stagnalis) gastropods.

Recent studies have highlighted that antidepressants such as the selective serotonin reuptake inhibitors (SSRIs) entering aquatic systems through wastewater discharges might impact organisms at environmentally relevant concentrations. In this study, two snail species (Gibbula unbilicalis and Lymnea stagnalis) representing the marine and freshwater environments were exposed to a large range of fluoxetine concentrations (1 ng L-1-1 mg L-1) and two distinct behaviours (foot detachment and righting time) were recorded. Fluoxetine significantly caused foot detachment only at the higher of the concentrations (1 mg L-1) in both species during the course of this short term 1.5 h and 4 h exposures. In this study, lowest observed effect concentrations (LOECs) for foot detachment fell repeatedly within the range for other gastropod snails exposed to fluoxetine. Fluoxetine effected righting times in a concentration dependant manner but only significantly within G. unbilicalis in the highest concentration. Reviewing existing data on the effects of antidepressants on a range of endpoints in gastropod molluscs reveals wide variability of results. The importance of publishing 'negative' and/or non-dramatic results to aid risk assessment are discussed along with the variability between antidepressants, model species, experimental designs and endpoints.

Impacts of a newly identified behaviour-altering trematode on its host amphipod: from the level of gene expression to population.

Changes to host behaviour induced by some trematode species, as a means of increased trophic transmission, represents one of the seminal examples of host manipulation by a parasite. The amphipod Echinogammarus marinus (Leach, 1815) is infected with a previously undescribed parasite, with infected individuals displaying positive phototaxic and negative geotaxic behaviour. This study reveals that the unknown parasite encysts in the brain, nerve cord and the body cavity of E. marinus, and belongs to the Microphallidae family. An 18 month population study revealed that host abundance significantly and negatively correlated with parasite prevalence. Investigation of the trematode's influence at the transcriptomic level revealed genes with putative neurological functions, such as serotonin receptor 1A, an inebriated-like neurotransmitter, tryptophan hydroxylase and amino acid decarboxylase, present consistent altered expression in infected animals. Therefore, this study provides one of the first transcriptomic insights into the neuronal gene pathways altered in amphipods infected with a trematode parasite associated with changes to its host's behaviour and population structure.

Crustacean intersexuality is feminization without demasculinization: implications for environmental toxicology.

The dysfunction associated with intersexuality in vertebrates and molluscs is often a serious threat to ecosystems. Although poorly understood, crustacean intersexuality is associated with contamination and includes forms linked to increased sex-ratio distorting parasites at polluted sites. Despite the importance of crustaceans for monitoring vulnerable aquatic habitats, little is known about the molecular basis of this abnormal sexual differentiation and any associated sexual dysfunction. To increase the relevance of crustaceans to environmental toxicologists, we comprehensively analyzed gene expression in amphipods presenting parasite- and nonparasite-associated intersexuality. Our findings reveal existing vertebrate biomarkers of feminization should not be applied to crustaceans, as orthologous genes are not induced in feminized amphipods. Furthermore, in contrast to vertebrates, where feminization and intersexuality is often associated with deleterious demasculinization, we find males maintain masculinity even when unambiguously feminized. This reveals a considerable regulatory separation of the gene pathways responsible for male and female characteristics and demonstrates that evidence of feminization (even if detected with appropriate biomarkers) is not a proxy for demasculinization in crustaceans. This study has also produced a comprehensive spectrum of potential molecular biomarkers that, when combined with our new molecular understanding, will greatly facilitate the use of crustaceans to monitor aquatic habitats.

Insights into PFAS contaminants before and after sewage discharges into a marine protected harbour.

Per and polyfluoroalkyl substances (PFAS) and their degradation products are a concern to human and ecosystem health. Wastewater treatment plants are not efficient at removing PFAS compounds and are thought to be a major source of these compounds to marine environments. The sewerage infrastructure in the UK, has over 20,000 combined stormwater overflows (CSOs). These CSOs are relief values whereby untreated wastewater can discharge under permit from the Environment Agency with exceptional rain/snowfall conditions. CSOs discharged 3.6 million monitored hours of untreated wastewater into English rivers and coasts in 2023. Concerns have been raised about the proximity of these CSO discharges to highly protected marine habitats. This study is the first to determine that PFAS concentrations are elevated in a highly protected marine bay (Langstone Harbour, England) following recent sewage releases compared to an extended period without discharge. Analysis was carried out into a suite of 54 PFAS compounds of which only one (PFHpA) was detectable above LOD prior to discharges but 8 afterwards. These included banned PFOS (Linear and Branched 8.6 ng/L ∓ 0.90) and PFOA (2.9 ng/L ∓ 0.29) which were above annual average EQS for inland and 'other' surface waters. Most of the PFAS compounds detected doubled in concentration above LODs. These two-fold increases we discuss are likely conservative estimates based on the use of LODs and tidal conditions. Additional Oysters (Crassostrea gigas) and Seaweed (Fucus vesiculosus) were taken revealing high concentrations of the shorter chain PFBA (6.99µg/kg ∓ 2.42 ww) in seaweed samples. These seaweeds were calculated to have conservative bioaccumulation factors (BAF) > 6000 for PFBA indicating these algae might be an important reservoir of some PFAS contamination. We discuss these results in the context of the largescale discharges of untreated wastewater nationally and globally, and call upon a need for a better understanding of the transfer of PFAS contaminants into marine food chains.

Evaluation of precopulatory pairing behaviour and male fertility in a marine amphipod exposed to plastic additives.

Plastics contain a mixture of chemical additives that can leach into the environment and potentially cause harmful effects on reproduction and the endocrine system. Two of these chemicals, N-butyl benzenesulfonamide (NBBS) and triphenyl phosphate (TPHP), are among the top 30 organic chemicals detected in surface and groundwater and are currently placed on international watchlist for evaluation. Although bans have been placed on legacy pollutants such as diethylhexyl phthalate (DEHP) and dibutyl phthalate (DBP), their persistence remains a concern. This study aimed to examine the impact of plastic additives, including NBBS, TPHP, DBP, and DEHP, on the reproductive behaviour and male fertility of the marine amphipod Echinogammarus marinus. Twenty precopulatory pairs of E. marinus were exposed to varying concentrations of the four test chemicals to assess their pairing behaviour. A high-throughput methodology was developed and optimised to record the contact time and re-pair time within 15 min and additional point observations for 96 h. The study found that low levels of NBBS, TPHP, and DEHP prolonged the contact and re-pairing time of amphipods and the proportion of pairs reduced drastically with re-pairing success ranging from 75% to 100% in the control group and 0%-85% in the exposed groups at 96 h. Sperm count declined by 40% and 60% in the 50 µg/l and 500 µg/l DBP groups, respectively, whereas TPHP resulted in significantly lower sperms in 50 µg/l exposed group. Animals exposed to NBBS and DEHP showed high interindividual variability in all exposed groups. Overall, this study provides evidence that plastic additives can disrupt the reproductive mechanisms and sperm counts of amphipods at environmentally relevant concentrations. Our research also demonstrated the usefulness of the precopulatory pairing mechanism as a sensitive endpoint in ecotoxicity assessments to proactively mitigate population-level effects in the aquatic environment.

The effects of plastic additives on swimming activity and startle response in marine amphipod Echinogammarus marinus.

Plastic additives are widely used in plastic production and are found in the environment owing to their widespread applications. Among these additives, N-butyl benzenesulfonamide (NBBS) and triphenyl phosphate (TPHP) are under international watchlist for evaluation, with limited studies on amphipods. Di-ethylhexyl phthalate (DEHP) and dibutyl phthalate (DBP) are banned in some countries and categorised as substances of very high concern. This study aimed to investigate the effects of NBBS, TPHP, DEHP and DBP on the swimming activity of a coastal intertidal marine amphipod, Echinogammarus marinus. Furthermore, this study is the first to quantify startle response in E. marinus in response to light stimuli. Amphipods were exposed to 0, 0.5, 5, 50 and 500 µg/l concentrations of all test compounds. Swimming activity and startle responses were assessed by video tracking and analysis using an 8-min alternating dark and light protocol after exposure on days 7 and 14. We observed an overall compound and light effect on the swimming activity of E. marinus. A significant decrease in swimming distance was found in 500 µg/l NBBS and TPHP. We observed that the startle response in E. marinus had a latency period of >2 s and animals were assessed at 1 s and the sum of the first 5 s. There was a clear startle response in E. marinus during dark to light transition, evident with increased swimming distance. NBBS exposure significantly increased startle response at environmental concentrations, while significant effects were only seen in 500 µg/l TPHP at 5 s. We found no significant effects of DEHP and DBP on swimming behaviour at the concentrations assessed. The findings of this study affirm the necessity for a continuous review of plastic additives to combat adverse behavioural effects that may be transferable to the population levels.

EthoCRED: a framework to guide reporting and evaluation of the relevance and reliability of behavioural ecotoxicity studies.

Behavioural analysis has been attracting significant attention as a broad indicator of sub-lethal toxicity and has secured a place as an important subdiscipline in ecotoxicology. Among the most notable characteristics of behavioural research, compared to other established approaches in sub-lethal ecotoxicology (e.g. reproductive and developmental bioassays), are the wide range of study designs being used and the diversity of endpoints considered. At the same time, environmental hazard and risk assessment, which underpins regulatory decisions to protect the environment from potentially harmful chemicals, often recommends that ecotoxicological data be produced following accepted and validated test guidelines. These guidelines typically do not address behavioural changes, meaning that these, often sensitive, effects are not represented in hazard and risk assessments. Here, we propose a new tool, the EthoCRED evaluation method, for assessing the relevance and reliability of behavioural ecotoxicity data, which considers the unique requirements and challenges encountered in this field. This method and accompanying reporting recommendations are designed to serve as an extension of the "Criteria for Reporting and Evaluating Ecotoxicity Data (CRED)" project. As such, EthoCRED can both accommodate the wide array of experimental design approaches seen in behavioural ecotoxicology, and could be readily implemented into regulatory frameworks as deemed appropriate by policy makers of different jurisdictions to allow better integration of knowledge gained from behavioural testing into environmental protection. Furthermore, through our reporting recommendations, we aim to improve the reporting of behavioural studies in the peer-reviewed literature, and thereby increase their usefulness to inform chemical regulation.

Radiocaesium accumulation and fluctuating asymmetry in the Japanese mitten crab, Eriocheir japonica, along a gradient of radionuclide contamination at Fukushima.

The 2011 Tohoku earthquake-tsunami and the subsequent nuclear accident at the Fukushima Dai-ichi Nuclear Power Station (FDNPS) led to large-scale radionuclide contamination of the marine and freshwater environment. Monitoring studies of marine food products in the Fukushima region have generally demonstrated a declining trend in radiocaesium concentrations. However, the accumulation and elimination of radiocaesium and potential biological effects remain poorly understood for freshwater biota inhabiting highly contaminated areas at Fukushima. Consequently, the present study aimed to assess radiocaesium accumulation and developmental effects on the commercially important catadromous Japanese mitten crab, Eriocheir japonica. E. japonica were collected from four sites along a gradient of radionuclide contamination 4-44 km in distance from the FDNPS in 2017. To determine potential developmental effects, fluctuating asymmetry (FA) was used as a measure of developmental stability. Combined 134Cs and 137Cs values for whole E. japonica from highly contaminated sites 4 and 16 km in distance from the FDNPS were 3040 ± 521 and 2250 ± 908 Bq kg-1 wet weight respectively, 30 and 22 times greater than the Japanese standard limit of 100 Bq kg-1. Estimated total dose rates based on radiocaesium concentrations in whole crabs and sediment ranged from 0.016 to 37.7 µGy h-1. No significant relationship between radiocaesium accumulation and FA was recorded, suggesting that chronic radiation exposure at Fukushima is not inducing developmental effects in E. japonica as measured using fluctuating asymmetry. Furthermore, estimated dose rates were below proposed regulatory limits where significant deleterious effects are expected. The present study will aid in the understanding of the long-term consequences of radiation exposure for non-human biota and the management of radioactively contaminated environments.

Shipping in the north-east Atlantic: Identifying spatial and temporal patterns of change.

Maritime traffic is increasing globally, with a four-fold increase in commercial vessel movements between 1992 and 2012. Vessels contribute to noise and air pollution, provide pathways for non-native species, and collide with marine wildlife. While knowledge of shipping trends and potential environmental impacts exists at both local and global levels, key information on vessel density for regional-scale management is lacking. This study presents the first in-depth spatio-temporal analysis of shipping in the north-east Atlantic region, over three years in a five-year period. Densities increased by 34%, including in 73% of Marine Protected Areas. Western Scotland and the Bay of Biscay experienced the largest increases in vessel density, predominantly from small and slow vessels. Given well-documented impacts that shipping can have on the marine environment, it is crucial that this situation continues to be monitored - particularly in areas designated to protect vulnerable species and ecosystems which may already be under pressure.

Biomimetic generation of the strongest known biomaterial found in limpet tooth.

The biomaterial with the highest known tensile strength is a unique composite of chitin and goethite (α-FeO(OH)) present in teeth from the Common Limpet (Patella vulgata). A biomimetic based on limpet tooth, with corresponding high-performance mechanical properties is highly desirable. Here we report on the replication of limpet tooth developmental processes ex vivo, where isolated limpet tissue and cells in culture generate new biomimetic structures. Transcriptomic analysis of each developmental stage of the radula, the organ from which limpet teeth originate, identifies sequential changes in expression of genes related to chitin and iron processing. We quantify iron and chitin metabolic processes in the radula and grow isolated radula cells in vitro. Bioinspired material can be developed with electrospun chitin mineralised by conditioned media from cultured radula cells. Our results inform molecular processes behind the generation of limpet tooth and establish a platform for development of a novel biomimetic with comparable properties.

Frontiers in quantifying wildlife behavioural responses to chemical pollution.

Animal behaviour is remarkably sensitive to disruption by chemical pollution, with widespread implications for ecological and evolutionary processes in contaminated wildlife populations. However, conventional approaches applied to study the impacts of chemical pollutants on wildlife behaviour seldom address the complexity of natural environments in which contamination occurs. The aim of this review is to guide the rapidly developing field of behavioural ecotoxicology towards increased environmental realism, ecological complexity, and mechanistic understanding. We identify research areas in ecology that to date have been largely overlooked within behavioural ecotoxicology but which promise to yield valuable insights, including within- and among-individual variation, social networks and collective behaviour, and multi-stressor interactions. Further, we feature methodological and technological innovations that enable the collection of data on pollutant-induced behavioural changes at an unprecedented resolution and scale in the laboratory and the field. In an era of rapid environmental change, there is an urgent need to advance our understanding of the real-world impacts of chemical pollution on wildlife behaviour. This review therefore provides a roadmap of the major outstanding questions in behavioural ecotoxicology and highlights the need for increased cross-talk with other disciplines in order to find the answers.

Priorities to inform research on marine plastic pollution in Southeast Asia.

Southeast Asia is considered to have some of the highest levels of marine plastic pollution in the world. It is therefore vitally important to increase our understanding of the impacts and risks of plastic pollution to marine ecosystems and the essential services they provide to support the development of mitigation measures in the region. An interdisciplinary, international network of experts (Australia, Indonesia, Ireland, Malaysia, the Philippines, Singapore, Thailand, the United Kingdom, and Vietnam) set a research agenda for marine plastic pollution in the region, synthesizing current knowledge and highlighting areas for further research in Southeast Asia. Using an inductive method, 21 research questions emerged under five non-predefined key themes, grouping them according to which: (1) characterise marine plastic pollution in Southeast Asia; (2) explore its movement and fate across the region; (3) describe the biological and chemical modifications marine plastic pollution undergoes; (4) detail its environmental, social, and economic impacts; and, finally, (5) target regional policies and possible solutions. Questions relating to these research priority areas highlight the importance of better understanding the fate of marine plastic pollution, its degradation, and the impacts and risks it can generate across communities and different ecosystem services. Knowledge of these aspects will help support actions which currently suffer from transboundary problems, lack of responsibility, and inaction to tackle the issue from its point source in the region. Being profoundly affected by marine plastic pollution, Southeast Asian countries provide an opportunity to test the effectiveness of innovative and socially inclusive changes in marine plastic governance, as well as both high and low-tech solutions, which can offer insights and actionable models to the rest of the world.